Novel use of pyrrolopyridine derivatives for preventing or treating inflammatory diseases

ABSTRACT

The present invention relates to a pharmaceutical composition for preventing and/or treating inflammatory disease, the composition comprising a pyrrolopyridine-derivative compound as an active ingredient; the compound represented by Chemical Formula 1 according to the present invention, enantiomers thereof or pharmaceutically acceptable salts thereof, which have excellent inhibitory activity against not only protein kinases but also inflammatory factors and inflammatory cytokines; therefore a pharmaceutical composition comprising the same as an active ingredient may be useful in prevention, treatment and/or improvement of inflammatory disease, in particular but not limited to inflammatory bowel disease, rheumatoid arthritis, or lupus.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a novel use of pyrrolopyridinederivative compounds for preventing and/or treating inflammatorydiseases.

BACKGROUND OF THE INVENTION

Pyrrolopyridine derivatives are bicyclic compounds wherein pyridine andpyrrole are bonded and used for various pharmaceutical purposes.Accordingly, various pyrrolopyridine derivatives have been synthesized.Patent literature which discloses pyrrolopyridine derivatives includeKorean Laid-open Patent Nos. 10-2018-0015142, 10-2017-0106452, and10-2017-0058465.

In most of the above patent literature, it can be known thatpyrrolopyridine derivatives are used as inhibitors for protein kinases.Accordingly, the inventors set out to identify other physiologicalactivities and pharmacological mechanisms of the pyrrolopyridinederivatives.

Inflammation is the mechanism by which living tissue that has beendamaged by physical action, chemical substance, or bacterial infectionis restored. When said stimuli are applied, vasoactive substances suchas histamine, serotonin, and prostaglandin are released, increasingvascular permeability and causing inflammation.

It has been reported that when macrophages are activated bylipopolysaccharides (LPS), the TLR (toll-like receptor)-4 signal pathwayincluding phosphorylation of mitogen-activated protein kinase (MAPK) andactivation of transcription factor NF-κB (nuclear factor-kappa B) isinitiated, which induces the generation of inflammation-related factorssuch as NO, IL-6, IL-1β, TNF-α, iNOS and COX-2 (Chae H S, et al. BiolPharm Bull. 2009; 32(4):553-557.).

Recently, there has been active research into methods of reducinginflammation generation, and nonsteroidal anti-inflammatory drugs(NSAIDs) including bradykinin antagonists and TNF-α inhibitors havinganti-inflammatory, analgesic, and antipyretic actions are commonly usedas anti-inflammatory drugs. However, long-term use of non-steroidalanti-inflammatory drugs may cause gastrointestinal disorders, andserious side effects such as secondary anemia, asthma, suppression oflabor induction, adverse effects on the kidneys, liver damage, andhypersensitivity have been reported.

Therefore, there is a need for the development of an anti-inflammatoryagent which minimizes the usual and often seen side effects of currentlyknown anti-inflammatory agents, does not harm the human body especiallyfor maintenance use, and can be administered over an extended period oftime.

Thereupon, the present inventors discovered that pyrrolopyridinederivatives inhibit the expression of inflammation-related factorsincluding inflammatory cytokines such as NO, IL-6, IL-1β, TNF-α and thelike, and by confirming in-vivo stability and safety of the same, haveidentified potential for the use of pyrrolopyridine derivatives aspharmaceutical agents for the prevention and/or treatment ofinflammatory diseases.

SUMMARY OF THE INVENTION

A purpose of the present invention is to provide a pharmaceuticalcomposition for prevention and/or treatment of inflammatory diseases,the composition comprising a pyrrolopyridine-derivative compound as anactive ingredient.

Another purpose of the present invention is to provide a method forpreventing and/or treating inflammatory diseases by administering aneffective dose of a pyrrolopyridine-derivative compound to a subject.

Yet another purpose of the present invention is to provide apyrrolopyridine-derivative composition for prevention and/or treatmentof inflammatory disease, and a use for a pharmaceutical compositioncomprising the same as an active ingredient.

In one embodiment, the present invention provides a method of preventingand/or treating an inflammatory disease in a patient in need thereof,the method comprising administering to the patient a compoundrepresented by Chemical Formula 1:

or an isomer, pharmaceutically acceptable salt, or pharmaceuticalcomposition thereof, wherein:R¹ is C₁-C₃ alkoxy;R² and R³ are each independently hydrogen, straight-chain or branchedC₁-C₁₀ alkyl, or C₃-C₆ cycloalkyl;

and

R⁴ is haloalkyl.

In one embodiment, the present invention provides a method forpreventing and/or treating inflammatory diseases, the method comprisinga step of administering an effective dose of a pharmaceuticalcomposition comprising a compound represented by Chemical Formula 1, oran isomer or pharmaceutically acceptable salt thereof.

The present invention provides a use of a compound represented byChemical Formula 1, an isomer thereof, a pharmaceutically acceptablesalt thereof, or a pharmaceutical composition comprising the same as anactive ingredient, for preventing and/or treating inflammatory diseases.

A compound represented by Chemical Formula 1 according to the presentinvention, or isomers or pharmaceutically acceptable salts thereof, hasexcellent inhibitory activity with regard to various protein kinasesincluding DYRK1A, CLK and the like, and/or excellent inhibitory activityagainst inflammation-related factors, while having excellent in-vivostability, and a pharmaceutical composition comprising the same as anactive ingredient can be useful in treating and/or preventinginflammatory diseases. In particular embodiments, a compound of ChemicalFormula 1 can be used for the prevention, treatment, and/or improvementof inflammatory bowel disease, psoriasis, rheumatoid arthritis, andlupus.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1a and FIG. 1b show inhibitory activity against expression of thepro-inflammatory cytokines TNF-α and IL-6 when LPS-stimulated THP-1cells are treated with Compound 1 (FIG. 1a ) and inhibitory activityagainst intranuclear mobility of p50 and p65 involved in the NF-Kbsignaling pathway (FIG. 1b ).

FIG. 2 shows the results of assessing regulation activity againstregulatory T cell and Th17 cell differentiation.

FIG. 3 shows the results of treating a collagen-induced arthritis mousemodel with Compound 1 to evaluate the therapeutic effects againstarthritis, showing results of arthritis index evaluation (a), jointweight measurement (b), and histologic analysis results (c) through (e)observed under a microscope after dyeing with H&E and T-blue.

FIG. 4 shows the results of evaluating the inhibitory activity ofCompound 1 on the secretion of inflammatory cytokines (TNF-α, IL-6 andIL-17A) when peripheral blood mononuclear cells in the blood ofinflammatory bowel disease patients have been treated with variousconcentrations.

FIG. 5 is a drawing of the result of optical microscope observationafter H/E dyeing the colon tissue of a TNBS-induced inflammatory boweldisease mouse model treated with Compound 1 and Enbrel and Cyclosporin Aas comparative compounds.

FIG. 6 shows the degree of expression of myeloperoxidase (MPO), as wellas the weight and histological analysis results of colon tissue aftertreatment of a 2,4,6-trinitrobenzenesulfonic acid (TNBS) induced colitismodel with Compound 1.

FIG. 7 shows the results of evaluating the efficacy after treating a DSS(dextran sodium sulfate)-induced inflammatory bowel disease mouse model,showing the results of evaluation weight loss suppression effect anddisease activity index (DAI).

FIG. 8a through FIG. 8c show the expression of inflammatory cytokines incolon tissue of a DSS-induced inflammatory bowel disease mouse modeltreated with Compound 1 and comparative compounds Cyclosporin A andTofacitinib (FIG. 8a ), an optical microscope photograph of colon tissueafter H/E dyeing (FIG. 8b ), and the results of scoring the weight tolength ratio and Goblet cell depletion of colon tissue (FIG. 8c ).

FIG. 9 shows the results of scoring the skin state of a house dustmite-induced atopic dermatitis model to analyze the therapeutic effectsof treating a house dust mite-induced atopic dermatitis model withCompound 1.

FIG. 10 shows the results of evaluating the inhibitory activity ofCompound 1 of IL-4, IL-6, MIP-1b, KC and IL-33 expression in skin tissueand IgE expression in blood serum when a house dust mite-induced atopicdermatitis model is administered with Compound 1 at varyingconcentrations.

FIG. 11a and FIG. 11b show the results of the topical anti-inflammatoryactivity of Compound 1 in the imiquimod (IMQ)-induced psoriasis-likeskin inflammation model.

FIG. 12a and FIG. 12b show the results of the oral anti-inflammatoryactivity of Compound 1 in the imiquimod (IMQ)-induced psoriasis-likeskin inflammation model.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS 1. General Description ofCertain Embodiments of the Invention

Examples of the present invention may be modified into various differentforms, and the scope of the present invention is not limited to theexamples described in the following. Furthermore, the examples of thepresent invention are provided to more completely describe the presentinvention to a person having ordinary skill in the art. Additionally, inthe entirety of the specification, “comprising” an element, unlessspecifically stated otherwise, does not exclude other elements and meansthat other elements may be further comprised.

2. Compounds and Definitions

Compounds of the present invention include those described generallyherein, and are further illustrated by the classes, subclasses, andspecies disclosed herein. As used herein, the following definitionsshall apply unless otherwise indicated. For purposes of this invention,the chemical elements are identified in accordance with the PeriodicTable of the Elements, CAS version, Handbook of Chemistry and Physics,75^(th) Ed. Additionally, general principles of organic chemistry aredescribed in “Organic Chemistry”, Thomas Sorrell, University ScienceBooks, Sausalito: 1999, and “March's Advanced Organic Chemistry”, 5^(th)Ed., Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001,the entire contents of which are hereby incorporated by reference.

As used herein, the term “administration” refers to using a suitablemethod to introduce the pharmaceutical composition of the presentinvention to a subject suspected to have inflammatory disease, andadministration may be performed through various pathways so long as thetarget tissue can be reached.

As used herein, the term “alkyl” may mean a straight-chain or branchedacyclic saturated hydrocarbon consisting of carbon atoms. Representative—(C₁₋₈alkyl) may include methyl, ethyl, n-propyl, n-butyl, n-pentyl,n-hexyl, n-heptyl and n-octyl; and branched chain saturated alkyls mayinclude isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl,2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, andthe like. The C₁₋₈ alkyl may be substituted or unsubstituted. Forexample, a C₁₋₈ alkyl group may be substituted with a phenyl to form abenzyl group.

As used herein, the phrase “conjointly administering” refers herein toany form of administration of two or more different therapeuticcompounds such that the second administered compound is administeredwhile the first administered therapeutic compound is still effective inthe body (e.g., the two compounds are simultaneously effective in thepatient, which may include additive or synergistic effects of the twocompounds).

As used herein, the term “cycloalkyl” may refer to a nonaromaticsaturated or unsaturated carbon ring. Representative cycloalkylsinclude, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclopentadienyl, cyclohexyl, cyclohexenyl, 1,3-cyclohexadienyl,1,4-cyclohexadienyl, cycloheptyl, 1,3-cycloheptadienyl,1,3,5-cycloheptatrienyl, cyclooctyl and cyclooctadienyl. The cycloalkylgroup may be substituted or unsubstituted. In one embodiment, thecycloalkyl group may be a C₃₋₈ cycloalkyl group. A cycloalkyl group ofC₇ or greater may have two or more cyclic structures, and a specificexample thereof may be a bicycloalkyl group, and more specifically,bicycloheptane may be used in the present invention.

At least one of the homogenous or heterogeneous substituents mentionedin the above may be substituted in like positions or differentpositions, and may also be sequentially substituted. The term“sequentially” in the above refers to one substituent being substitutedin a chemical formula followed by the consecutive substitution ofanother substituent, and, for example, in a case wherein an alkyl groupis substituted, and then a cycloalkyl is substituted at the alkyl group,and a carbonyl group is sequentially substituted at the cycloalkylgroup, the compound may be named carbonylcycloalkylalkyl to indicate ithas been sequentially substituted.

As used herein, the term “halogen” may be F, Cl, Br or I.

As used herein, the term “haloalkyl” may refer to a straight chain orbranched chain alkyl (hydrocarbon) having a carbon atom substituted byat least one halogen atom. Examples of haloalkyl include, but are notlimited to, methyl, ethyl, propyl, isopropyl, isobutyl and N-butylindependently substituted by at least one halogen atom, for example, F,Cl, Br or I.

As used herein, the term “hydrate” refers to a compound of the presentinvention or a salt thereof, comprising a stoichiometric ornon-stoichiometric amount of water bonded by a non-covalentintermolecular force. The hydrate of the compound represented by Formula1 of the present invention may include a stoichiometric ornon-stoichiometric amount of water that is bound by non-covalentintermolecular forces. Such hydrates may comprise at least oneequivalent, and preferably 1 to 5 equivalents, of water. Such hydratesmay be prepared by crystallizing, from water or a solvent comprisingwater, a compound represented by Chemical Formula 1 of the presentinvention, an isomer thereof, or pharmaceutically acceptable saltsthereof.

As used herein, the term “isomer” refers to a compound of the presentinvention or a salt thereof that has the same chemical formula ormolecular formula but differs structurally or sterically. Included insuch isomers are all structural isomers such as tautomers, R or Sisomers, stereoisomers such as geometrical isomers (trans, cis), andenantiomers. All such isomers and compounds thereof are too included inthe scope of the present invention. Unless otherwise specified, a solidline bond (—) connected to an asymmetric carbon atom may include a solidwedge bond

or dashed wedge bond

representing the absolute arrangement of a stereogenic center.

As used herein, the term “inflammatory disease” is a term generallyreferring to diseases whose principal condition is inflammation, andspecifically, but not limited to, may be one selected from asthma, acutelung injury, GvHD Graft versus Host Disease), chronic obstructivepulmonary disease, allergy, systemic lupus erythematosus, scleroderma,inflammatory bowel disease (for example ulcerative colitis and Crohn'sdisease), atopic dermatitis, psoriasis, anaphylaxis, dermatitis,diabetic retinosis, retinitis, macular degeneration, uveitis,conjunctivitis, rheumatoid arthritis, ankylosing spondylitis,osteoarthritis, osteoporosis, diabetes, diabetic kidney disease,nephritis, Sjogren's syndrome, Crohn's disease, autoimmune pancreatitis,periodontal disease, alopecia areata, graft versus host disease, chronicpelvic inflammatory disease, endometriosis, rhinitis, tonsilitis, otitismedia, sore throat, cystitis and chronic prostatitis. Furtherinflammatory diseases or diseases having an inflammatory component aredisclosed herein.

As used herein, the term “pharmaceutically effective dose” refers to adose sufficient to treat disease with a reasonable benefit/risk ratioapplicable to medical treatment, and the level of the effective dose maybe determined depending on factors including the type of subject,severity of disease, age, sex, type of disease, activity of the drug,drug sensitivity, duration of administration, administration pathway andexcretion rate, duration of therapy and concomitantly used drugs, andother factors known well to the medical art. The compounds orcompositions of the present invention may be administered as a singledrug or concomitantly with other drugs, such as by conjointadministration, and may be administered sequentially or simultaneouslywith commercially marketed therapeutic agents. Furthermore, it may beadministered as a single dose or multiple doses. It is important toadminister a dose capable of achieving the maximum effect with a minimumamount without developing adverse effects in consideration of all of theabove factors; the dose may be readily decided by a person of skill inthe art. The administration dose of the compound or composition of thepresent invention may be determined by an expert according to variousfactors such as patient status, age, sex and complications. As thecompound or composition of the present invention has excellent safety,it may be used at above the decided administration dose.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge etal., describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein byreference. Pharmaceutically acceptable salts of the compounds of thisinvention include those derived from suitable inorganic and organicacids and bases. Examples of pharmaceutically acceptable, nontoxic acidaddition salts are salts of an amino group formed with inorganic acidssuch as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuricacid and perchloric acid or with organic acids such as acetic acid,oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid ormalonic acid or by using other methods used in the art such as ionexchange. Other pharmaceutically acceptable salts include adipate,alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate,propionate, stearate, succinate, sulfate, tartrate, thiocyanate,p-toluenesulfonate, undecanoate, valerate salts, and the like.

Salts derived from appropriate bases include alkali metal, alkalineearth metal, ammonium and N⁺(C₁₋₄ alkyl)₄ salts. Representative alkalior alkaline earth metal salts include sodium, lithium, potassium,calcium, magnesium, and the like. Further pharmaceutically acceptablesalts include, when appropriate, nontoxic ammonium, quaternary ammonium,and amine cations formed using counterions such as halide, hydroxide,carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and arylsulfonate. In some embodiments, the provided compounds are purified insalt form for convenience and/or ease of purification, e.g., using anacidic or basic mobile phase during chromatography. Salts forms of theprovided compounds formed during chromotagraphic purification arecontemplated herein (e.g., diammonium salts) and are readily apparent tothose having skill in the art.

As used herein, the term “solvate” refers to a compound of the presentinvention or a salt thereof comprising a stoichiometric ornon-stoichiometric amount of solvent bonded by a non-covalentintermolecular force. Preferable solvents, therefore, include volatileor non-toxic solvents, and/or solvents suitable for humanadministration.

As used herein, the term “treatment” or “therapeutic” includes thesuppression, delay, identification, alleviation, weakening, limiting,reduction, inhibition, avoidance or healing of illnesses, conditions,disabilities, damage or health problems, or the occurrence orprogression/improvement of such statuses and/or symptoms of suchstatuses. The term “prevention” refers to avoidance or reduction of riskof contracting, experiencing, suffering or having illnesses, conditions,disabilities, damage or health problems, or the occurrence orprogression of such statuses and/or symptoms of such statuses. Treatmentor prevention of a disease, condition, disorder, injury or healthproblem may be partial or complete.

3. Description of Exemplary Embodiments

The present invention provides a compound for the prevention and/ortreatment of inflammatory diseases, the compound represented by ChemicalFormula 1:

or an isomer, pharmaceutically acceptable salt, or pharmaceuticalcomposition thereof, wherein:

R¹ is C₁-C₃ alkoxy;R² and R³ are each independently hydrogen, straight-chain or branchedC₁-C₁₀ alkyl, or C₃-C₆ cycloalkyl;

and

R⁴ is haloalkyl.

As defined and described herein, R¹ is C₁-C₃ alkoxy.

In some embodiments, R¹ is C₁-C₃ alkoxy. In some embodiments, R¹ ismethoxy.

In some embodiments, R¹ is as found in the compounds of Table 1, below.

As defined and described herein, R² and R³ are each independentlyhydrogen, straight-chain or branched C₁-C₁₀ alkyl, or C₃-C₆ cycloalkyl.

In some embodiments, R² is hydrogen. In some embodiments, R² is astraight-chain C₁-C₁₀ alkyl.

In some embodiments, R² is a branched C₁-C₁₀ alkyl. In some embodiments,R² is a C₃-C₆ cycloalkyl. In some embodiments, R² is straight chain orbranched chain C₁-C₅ alkyl or C₃-C₄ cycloalkyl.

In some embodiments, R³ is hydrogen. In some embodiments, R³ is astraight-chain C₁-C₁₀ alkyl.

In some embodiments, R³ is a branched C₁-C₁₀ alkyl. In some embodiments,R³ is a C₃-C₆ cycloalkyl.

In some embodiments, R² and R³ are as found in the compounds of Table 1,below.

As defined and described herein, R⁴ is haloalkyl.

In some embodiments, R⁴ is haloalkyl. In some embodiments, R⁴ istrifluoromethyl.

In certain embodiments, R¹ is methoxy; R² is straight chain or branchedchain C₁-C₅ alkyl or C₃-C₄ cycloalkyl; R³ is hydrogen; and R⁴ istrifluoromethyl.

In certain embodiments, a compound represented by Chemical Formula 1 is(4-((−4-ethylamino)-3-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-6-yl)amino)-3-methoxyphenyl)(4-morpholinopiperidine-1-yl)methanone.

In some embodiments, a compound represented by Chemical Formula 1 isfound in Table 1, below.

In some embodiments, a pharmaceutically acceptable salt of a compoundrepresented by Chemical Formula 1 may be interpreted as existing in anyform selected from a group comprising any crystalline or amorphous form,or hydrates, solvates or co-crystals thereof.

In some embodiments, a compound of present invention, a pharmaceuticallyacceptable salt, or a pharmaceutical composition thereof may act as aninhibitor for protein kinase. Furthermore, the compounds mentionedherein may act as inhibitors against inflammation-related factors (e.g.,cytokines).

In some embodiments, the protein kinase may be one or more of ALK, ALK(C1156Y), ALK (L1196M), CLK1, CLK2, CLK3, CLK4, CSNK1D, DYRK1A, DYRK1B,DYRK2, GAK, JNK1, LRRK2 (G2019S), LTK, MYLK, PAK2, PHKG1, PHKG2, STK33,ABL1-nonphosphorylated, CAMK2D, CAMKK2, CHEK2, CSNK1A1, CSNK1E, ERK5,HUNK, INSR, JAK1 (JH2domain-pseudokinase), JNK2, JNK3, LRRK2, MAPKAPK2,PLK4, and STK39.

Inflammation-related factors may refer to all factors known to bedirectly or indirectly associated with inflammatory reaction. In aspecific example of the present invention, inflammation-related factorsmay include pro-inflammatory cytokines, and inflammation-related factorswhose activity is inhibited by the compound of the present invention andthereby exhibit preventive and/or therapeutic effects againstinflammation may be, for example, selected from one or more of NO,IFN-γ, IL-1α, IL-1β, IL-4, IL-6, IL-10, IL-12, IL-13, IL-17, IL-17A,IL-22, IL-23, IL-33, KC, MIP-1a, MIP-1b, GM-CSF, and TNF-α.

In some embodiments, the present invention provides a method of reducingpro-inflammatory cytokines in a patient in need thereof, the methodcomprising administering an effective dose of a compound represented byChemical Formula 1 or an isomer, pharmaceutically acceptable salt, or apharmaceutical composition thereof to a patient. In some embodiments,the method of reducing pro-inflammatory cytokines reduces one or more ofNO, IFN-γ, IL-1α, IL-1β, IL-4, IL-6, IL-10, IL-12, IL-13, IL-17, IL-17A,IL-22, IL-23, IL-33, KC, MIP-1α, MIP-1α, GM-CSF, and TNF-α. In certainembodiments, the method of reducing pro-inflammatory cytokines reducesone or more of IL-6, IL-17A, and TNR-α. In certain embodiments, themethod of reducing pro-inflammatory cytokines reduces one or more ofIL-4, IL-6, MIP-1b, KC and IL-33. In some embodiments, the disclosedmethods reduce IL-4. In some embodiments, the disclosed methods reduceIL-6. In some embodiments, the disclosed methods reduce IL-17A. In someembodiments, the disclosed methods reduce TNR-α. In some embodiments,the disclosed methods reduce MIP-1b. In some embodiments, the disclosedmethods reduce KC. In some embodiments, the disclosed methods reduceIL-33. In some embodiments, the method of decreasing pro-inflammatorycytokines in a patient comprises preventing and/or treating inflammatorydiseases, such as, but not limited to, inflammatory bowel disease orcolitis.

In some embodiments, the present invention provides a method of reducingIgE in a patient in need thereof, the method comprising administering aneffective dose of a compound represented by Chemical Formula 1, or anisomer, pharmaceutically acceptable salt, or a pharmaceuticalcomposition thereof to the patient.

In some embodiments, the present invention provides a method ofincreasing differentiation of regulatory T cells (T_(reg)) in a patientin need thereof, the method comprising administering an effective doseof a compound represented by Chemical Formula 1, or an isomer,pharmaceutically acceptable salt, or a pharmaceutical compositionthereof to a patient. In some embodiments, the method of increasingdifferentiation of regulatory T cells (T_(reg)) in a patient comprisespreventing and/or treating inflammatory diseases, such as, but notlimited to, an autoimmune disease.

In some embodiments, the present invention provides a method ofdecreasing pro-inflammatory T cells (e.g., Th17) in a patient in needthereof, the method comprising administering an effective dose of acompound represented by Chemical Formula 1, or an isomer,pharmaceutically acceptable salt, or a pharmaceutical compositionthereof to a patient. In some embodiments, the method of decreasingpro-inflammatory T cells (e.g., Th17) in a patient comprises preventingand/or treating inflammatory diseases, such as, but not limited to, anautoimmune disease.

In some embodiments, the present invention provides a method ofincreasing differentiation of regulatory T cells (T_(reg)) anddecreasing pro-inflammatory T cells (e.g., Th17) in a patient in needthereof, the method comprising administering an effective dose of acompound represented by Chemical Formula 1, or an isomer,pharmaceutically acceptable salt, or a pharmaceutical compositionthereof to a patient. In some embodiments, the method of increasingdifferentiation of regulatory T cells (T_(reg)) and decreasingpro-inflammatory T cells (e.g., Th17) in a patient comprises preventingand/or treating inflammatory diseases, such as, but not limited to, anautoimmune disease.

In some embodiments the inflammatory disease which can be treatedaccording to the methods of this invention is a Th17-mediated disease,but is not limited to that disease. In some embodiments theTh17-mediated disease is selected from systemic lupus erythematosus,multiple sclerosis, and/or an inflammatory bowel disease, such as, butnot limited to, Crohn's disease or ulcerative colitis.

In some embodiments, the present invention provides a method forpreventing and/or treating inflammatory diseases for a patient in needthereof, the method comprising administering an effective dose of acompound represented by Chemical Formula 1, or an isomer,pharmaceutically acceptable salt, or a pharmaceutical compositionthereof to the patient.

In some embodiments, a compound of the present invention represented byChemical Formula 1, or an isomer, pharmaceutically acceptable salt, or apharmaceutical composition thereof are useful in the prevention and/ortreatment of inflammatory or obstructive airway diseases, resulting, forexample, but not limited to, in reduction of tissue damage, airwayinflammation, bronchial hyperreactivity, remodeling, or diseaseprogression. In some embodiments, obstructive airway diseases includeobstructive airway diseases having an inflammatory component.Inflammatory or obstructive airway diseases to which the presentinvention is applicable include but are not limited to: asthma ofwhatever type or genesis including both intrinsic (non-allergic) asthmaand extrinsic (allergic) asthma, mild asthma, moderate asthma, severeasthma, bronchitic asthma, exercise-induced asthma, occupational asthmaand asthma induced following bacterial infection. Treatment of asthmaalso is to be understood as embracing treatment of subjects, e.g., ofless than 4 or 5 years of age, exhibiting wheezing symptoms anddiagnosed or diagnosable as “wheezy infants”, an established patientcategory of major medical concern and now often identified as incipientor early-phase asthmatics.

In some embodiments, a compound of the present invention represented byChemical Formula 1, or an isomer, pharmaceutically acceptable salt, or apharmaceutical composition thereof are useful in the prevention and/ortreatment of heteroimmune diseases. Examples of such heteroimmunediseases include, but are not limited to, graft versus host disease,transplantation, transfusion, anaphylaxis, allergies (e.g., allergies toplant pollens, latex, drugs, foods, insect poisons, animal hair, animaldander, dust mites, cockroach calyx, or other sources), type Ihypersensitivity, allergic conjunctivitis, allergic rhinitis, and atopicdermatitis.

Prophylactic efficacy in the treatment of asthma will be evidenced forthis invention by reduced frequency or severity of symptomatic attack,but not limited to this, e.g., of acute asthmatic or bronchoconstrictorattack, improvement in lung function, and/or improved airwayhyperreactivity. Such efficacy may further be evidenced by reducedrequirement for other symptomatic therapy, such as, but not limited to,therapy for or intended to restrict or abort symptomatic attack when itoccurs, for example anti-inflammatory or bronchodilatory. Prophylacticbenefit in asthma may in particular be apparent in subjects prone to“morning dipping”. Morning dipping is a recognized asthmatic syndrome,common to a substantial percentage of asthmatics and characterized byasthma attack, e.g., between the hours of about 4 to 6 AM, i.e., at atime normally substantially distant from any previously administeredsymptomatic asthma therapy.

In some embodiments, a compound of the present invention represented byChemical Formula 1, or an isomer, pharmaceutically acceptable salt, or apharmaceutical composition thereof are useful in the prevention and/ortreatment of other inflammatory or obstructive airway diseases andconditions to which the present invention is applicable and include butare not limited to acute lung injury (ALI), adult/acute respiratorydistress syndrome (ARDS), chronic obstructive pulmonary, airways or lungdisease (COPD, COAD or COLD), including chronic bronchitis or dyspneaassociated therewith, emphysema, as well as exacerbation of airwayhyperreactivity consequent to other drug therapy, in particular otherinhaled drug therapy. The invention also is applicable to the preventionand/or treatment of bronchitis of whatever type or genesis including,but not limited to, acute, arachidic, catarrhal, croupous, chronic, orphthinoid bronchitis. Further inflammatory or obstructive airwaydiseases to which the present invention is applicable include but arenot limited to pneumoconiosis (an inflammatory, commonly occupational,disease of the lungs, frequently accompanied by airway obstruction,whether chronic or acute, and occasioned by repeated inhalation ofdusts) of whatever type or genesis, including, for example, aluminosis,anthracosis, asbestosis, chalicosis, ptosis, siderosis, silicosis,tabacosis, and/or byssinosis.

With regard to their anti-inflammatory activity, in particular inrelation to inhibition of eosinophil activation, a compound of thepresent invention represented by Chemical Formula 1, or an isomer,pharmaceutically acceptable salt, or a pharmaceutical compositionthereof are also useful in the prevention and/or treatment ofeosinophil-related disorders, e.g. eosinophilia, in particulareosinophil-related disorders of the airways (e.g., involving morbideosinophilic infiltration of pulmonary tissues) including but notlimited to hypereosinophilia as it affects the airways and/or lungs aswell as, for example, eosinophil-related disorders of the airwaysconsequential or concomitant to Loffler's syndrome, eosinophilicpneumonia, parasitic (in particular metazoan) infestation (includingtropical eosinophilia), bronchopulmonary aspergillosis, polyarteritisnodosa (including Churg-Strauss syndrome), eosinophilic granuloma,and/or eosinophil-related disorders affecting the airways occasioned bydrug reaction.

In some embodiments, a compound of the present invention represented byChemical Formula 1, or an isomer, pharmaceutically acceptable salt, or apharmaceutical composition thereof are useful in the prevention and/ortreatment of inflammatory or allergic conditions of the skin; forexample, but not limited to, psoriasis, contact dermatitis, atopicdermatitis, alopecia areata, erythema multiforme, dermatitisherpetiformis, hidradenitis suppurativa, scleroderma, vitiligo,hypersensitivity angiitis, urticaria, bullous pemphigoid, lupuserythematosus, systemic lupus erythematosus, pemphigus vulgaris,pemphigus foliaceus, paraneoplastic pemphigus, epidermolysis bullosaacquisita (EBA), acne vulgaris, and/or other inflammatory or allergicconditions of the skin.

In some embodiments, a compound of the present invention represented byChemical Formula 1, or an isomer, pharmaceutically acceptable salt, or apharmaceutical composition thereof are useful for the prevention and/ortreatment of inflammatory diseases or conditions, including but notlimited to diseases or conditions having an inflammatory component; forexample, graft versus host disease (GvHD), chronic obstructive airwaydisease, obstructive pulmonary disease, allergy, systemic lupuserythematosus, scleroderma, inflammatory bowel disease (e.g., ulcerativecolitis and Crohn's disease), atopic dermatitis, psoriasis, anaphylaxis,dermatitis, diabetic retinosis, retinitis, macular degeneration,uveitis, conjunctivitis, rheumatoid arthritis, ankylosing spondylitis,osteoarthritis, osteoporosis, diabetes, diabetic kidney disease,nephritis, Sjogren's syndrome, autoimmune pancreatitis, periodontaldisease, alopecia areata, chronic pelvic inflammatory disease,endometriosis, rhinitis, tonsilitis, otitis media, sore throat,cystitis, chronic prostatitis, diseases and conditions of the eye suchas ocular allergy, keratoconjunctivitis sicca, vernal conjunctivitis,diseases affecting the nose including allergic rhinitis, and/orinflammatory disease in which autoimmune reactions are implicated orhaving an autoimmune component or etiology, including but not limited toautoimmune hematological disorders (e.g., hemolytic anemia, aplasticanemia, pure red cell anemia and idiopathic thrombocytopenia),polychondritis, Wegener granulomatosis, dermatomyositis, chronic activehepatitis, myasthenia gravis, Steven-Johnson syndrome, idiopathic sprue,irritable bowel syndrome, celiac disease, periodontitis, hyalinemembrane disease, kidney disease, glomerular disease, alcoholic liverdisease, multiple sclerosis, endocrine ophthalmopathy, Becker's/Duchennemuscular dystrophy, Grave's disease, sarcoidosis, alveolitis, chronichypersensitivity pneumonitis, multiple sclerosis, primary biliarycirrhosis, uveitis (anterior and posterior), interstitial lung fibrosis,psoriatic arthritis, systemic juvenile idiopathic arthritis,cryopyrin-associated periodic syndrome, vasculitis, diverticulitis,interstitial cystitis, glomerulonephritis (with and without nephroticsyndrome, e.g. including idiopathic nephrotic syndrome or minimal changenephropathy), chronic granulomatous disease, leptospirosis renaldisease, glaucoma, retinal disease, aging, headache, pain, complexregional pain syndrome, cardiac hypertrophy, muscle atrophy, catabolicdisorders, obesity, fetal growth retardation, hypercholesterolemia,heart disease, chronic heart failure, mesothelioma, anhidroticectodermal dysplasia, Behcet's disease, incontinentia pigmenti, Paget'sdisease, pancreatitis, hereditary periodic fever syndrome, asthma(allergic and non-allergic, mild, moderate, severe, bronchitic, andexercise-induced), acute lung injury, acute respiratory distresssyndrome, eosinophilia, hypersensitivities, nasal sinusitis, ocularallergy, silica induced diseases, COPD (reduction of damage, airwaysinflammation, bronchial hyperreactivity, remodeling or diseaseprogression), pulmonary disease, cystic fibrosis, acid-induced lunginjury, pulmonary hypertension, polyneuropathy, cataracts, muscleinflammation in conjunction with systemic sclerosis, inclusion bodymyositis, myasthenia gravis, thyroiditis, Addison's disease, lichenplanus, diabetes (Type 1 diabetes or Type 2 diabetes), appendicitis,blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis,cholangitis, cholecystitis, chronic graft rejection, colitis,dacryoadenitis, dermatomyositis, encephalitis, endocarditis,endometritis, enteritis, enterocolitis, epicondylitis, epididymitis,fasciitis, fibrositis, gastritis, gastroenteritis, Henoch-Schonleinpurpura, hepatitis, hidradenitis suppurativa, immunoglobulin Anephropathy, interstitial lung disease, laryngitis, mastitis,meningitis, myelitis myocarditis, myositis, nephritis, oophoritis,orchitis, osteitis, otitis, parotitis, pericarditis, peritonitis,pharyngitis, pleuritis, phlebitis, pneumonitis, pneumonia, polymyositis,proctitis, prostatitis, pyelonephritis, salpingitis, sinusitis,stomatitis, synovitis, tendonitis, tonsillitis, ulcerative colitis,vaginitis, vasculitis, and/or vulvitis.

In some embodiments, the inflammatory disease is, but not limited to,asthma, acute lung injury, GvHD Graft versus Host Disease), chronicobstructive pulmonary disease, allergy, systemic lupus erythematosus,scleroderma, inflammatory bowel disease (for example ulcerative colitisand Crohn's disease), atopic dermatitis, psoriasis, anaphylaxis,dermatitis, diabetic retinosis, retinitis, macular degeneration,uveitis, conjunctivitis, rheumatoid arthritis, ankylosing spondylitis,osteoarthritis, osteoporosis, diabetes, diabetic kidney disease,nephritis, Sjogren's syndrome, Crohn's disease, autoimmune pancreatitis,periodontal disease, alopecia areata, graft versus host disease, chronicpelvic inflammatory disease, endometriosis, rhinitis, tonsilitis, otitismedia, sore throat, cystitis and chronic prostatitis. In certainembodiments, the inflammatory disease is, but not limited to,inflammatory bowel disease, psoriasis, rheumatoid arthritis and lupus.

In some embodiments, the inflammatory disease which can be preventedand/or treated according to the methods of this invention is but is notlimited to a disease of the skin. In some embodiments, the inflammatorydisease of the skin is but is not limited to being selected from contactdermatitis, atopic dermatitis, alopecia areata, erythema multiforme,dermatitis herpetiformis, scleroderma, vitiligo, hypersensitivityangiitis, urticaria, bullous pemphigoid, pemphigus vulgaris,pemphigusfoliaceus, paraneoplastic pemphigus, epidermolysis bullosaacquisita (EBA), and/or other inflammatory or allergic conditions of theskin.

In some embodiments, the inflammatory disease which can be preventedand/or treated according to the methods of this invention is but is notlimited to being selected from acute and chronic gout, chronic goutyarthritis, psoriasis, psoriatic arthritis, rheumatoid arthritis,juvenile rheumatoid arthritis, systemic juvenile idiopathic arthritis(SEA), cryopyrin associated periodic syndrome (CAPS), andosteoarthritis.

In some embodiments, the inflammatory disease which can be preventedand/or treated according to the methods of this invention is but is notlimited to being selected from Sjogren's syndrome, allergic disorders,osteoarthritis, conditions of the eye such as ocular allergy,conjunctivitis, keratoconjunctivitis sicca and vernal conjunctivitis,and/or diseases affecting the nose, such as allergic rhinitis.

Furthermore, the present invention provides a method for improving,preventing, treating and/or alleviating inflammation occurring on theskin and/or mucous membranes, the method comprising applying, on apatient requiring improvement and/or alleviation of inflammationoccurring on the skin and/or mucous membranes, a compound represented byChemical Formula 1, or an isomer or pharmaceutically acceptable saltthereof, or a pharmaceutical composition comprising the same.

Furthermore, the present invention provides a use of a compoundrepresented by Chemical Formula 1, an isomer thereof, or apharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition thereof, for use in preventing and/or treating inflammatorydiseases. Specifically, the present invention provides a use of acompound represented by Chemical Formula 1, an isomer thereof, apharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition thereof, for the production of medicaments for use inpreventing and/or treating inflammatory diseases.

The pharmaceutical composition of the present invention, by furthercomprising excipients, disintegrating agents, sweetening agents,lubricants, flavoring agents and the like, may be formulated usingordinary methods into tablets, capsules, powders, granules, suspensions,emulsions, syrups, topicals, or other liquid or semi-liquid or solidformulations. For example, as the pharmaceutical composition of thepresent invention may act systemically and/or locally, and may beadministered to subjects orally or non-orally, that is, through variouspathways such as but not limited to pulmonary route administration,intranasal administration, sublingual administration, lingualadministration, buccolingual administration, rectal administration,dermal administration, transdermal administration, or conjunctivaladministration, the pharmaceutical composition may be formulated into aform suitable for the administration pathway. For example, dosage formssuitable for oral administration are but not limited to formulationscomprising a compound of the present invention in crystalline and/oramorphous and/or dissolved form, and may be, for example, tablets(coated or non-coated tablets, for example, using gastricfluid-resistant, delayed-dissolution or insoluble coatings), tablets orfilms/oblates which rapidly disintegrate in the oral cavity,films/freeze-dried preparations, capsules (for example, hard or softgelatin capsules), sugar-coated tablets, chewables (for example, softchewables), granules, pellets, powders, emulsions, suspensions, aerosolsand/or solutions. Parenteral administration may be achieved by avoidingthe absorption stage (for example, through intravenous, intraarterial,intra cardiac, intrathecal or intra lumbar administration) or includingabsorption (for example, through intramuscular, dermal, intradermal,subdermal, percutaneous or intraperitoneal pathways). Dosage formssuitable for parenteral administration may include but are not limitedto solutions, suspensions, emulsions, freeze-dried preparations, orpreparations in the form of sterilized powders for injection.

In some embodiments, the present invention provides a method forpreventing and/or treating inflammatory diseases for a patient in needthereof, the method comprising conjointly administering an effectivedose of a compound represented by Chemical Formula 1, or an isomer,pharmaceutically acceptable salt, or pharmaceutical composition thereofand at least one additional active ingredient.

In some embodiments, the pharmaceutical composition of the presentinvention may, for more effective treatment and/or prevention ofinflammatory disease, comprise, in addition to a compound of the presentinvention, at least one additional active ingredient. Active ingredientsthat may be suitable for combination include but are not limited togamma globulin, immunomodulatory and immunosuppressive compounds (forexample, cyclosporine, Methotrexate®), TNF antagonists (for example,Humira®, etanercept, infliximab), IL-1 inhibitors (for example,anakinra, canakinumab, rilonacept), phosphodiesterase inhibitors (forexample, apremilast), JAK/STAT inhibitors (for example, tofacitinib,baricitinib, GLPG0634), IRAK4 inhibitors, leflunomide, cyclophosphamide,rituximab, belimumab, tacrolimus, rapamycin, mycophenolate mofetil,interferon, corticosteroids (for example, prednisone, prednisolone,methylprednisolone, hydrocortisone, betamethasone), cyclophosphamide,azathioprine, sulfasalazine, paracetamol, and/or a non-steroidalanti-inflammatory agent (NSAID) (for example, aspirin, ibuprofen,naproxen, etodolac, celecoxib, colchicine).

Additional active agents that may be used in combination with a compoundrepresented by Chemical Formula 1, or an isomer, pharmaceuticallyacceptable salt, or pharmaceutical composition thereof, may be but arenot limited to small molecules or recombinant biologic agents andinclude, for example, acetaminophen, non-steroidal anti-inflammatorydrugs (NSAIDS) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®)and celecoxib, colchicine (Colcrys®), corticosteroids such asprednisone, prednisolone, methylprednisolone, hydrocortisone, and thelike, probenecid, allopurinol, febuxostat (Uloric®), sulfasalazine(Azulfidine®), antimalarials such as hydroxychloroquine (Plaquenil®) andchloroquine (Aralen®), methotrexate (Rheumatrex®), gold salts such asgold thioglucose (Solganal®), gold thiomalate (Myochrysine®) andauranofin (Ridaura®), D-penicillamine (Depen® or Cuprimine®),azathioprine (Imuran®), cyclophosphamide (Cytoxan®), chlorambucil(Leukeran®), cyclosporine (Sandimmune®), leflunomide (Araya®) and“anti-TNF” agents such as etanercept (Enbrel®), infliximab (Remicade®),golimumab (Simponi®), certolizumab pegol (Cimzia®) and adalimumab(Humira®), “anti-IL-1” agents such as anakinra (Kineret®) and rilonacept(Arcalyst®), canakinumab (Ilaris®), anti-Jak inhibitors such astofacitinib, antibodies such as rituximab (Rituxan®), “anti-T-cell”agents such as abatacept (Orencia®), “anti-IL-6” agents such astocilizumab (Actemra®), diclofenac, cortisone, hyaluronic acid (Synvisc®or Hyalgan®), monoclonal antibodies such as tanezumab, anticoagulantssuch as heparin (Calcinparine® or Liquaemin®) and warfarin (Coumadin®),antidiarrheals such as diphenoxylate (Lomotil®) and loperamide(Imodium®), bile acid binding agents such as cholestyramine, alosetron(Lotronex®), lubiprostone (Amitiza®), laxatives such as Milk ofMagnesia, polyethylene glycol (MiraLax®), Dulcolax®, Correctol® andSenokot®, anticholinergics or antispasmodics such as dicyclomine(Bentyl®), Singulair®, beta-2 agonists such as albuterol (Ventolin® HFA,Proventil® HFA), levalbuterol (Xopenex®), metaproterenol (Alupent®),pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®),salmeterol xinafoate (Serevent®) and formoterol (Foradil®),anticholinergic agents such as ipratropium bromide (Atrovent®) andtiotropium (Spiriva®), inhaled corticosteroids such as beclomethasonedipropionate (Beclovent®, Qvar®, and Vanceril®), triamcinolone acetonide(Azmacort®), mometasone (Asthmanex®), budesonide (Pulmocort®), andflunisolide (Aerobid®), Afviar®, Symbicort®, Dulera®, cromolyn sodium(Intal®), methylxanthines such as theophylline (Theo-Dur®, Theolair®,Slo-Bid®, Uniphyl®, Theo-24®) and aminophylline, IgE antibodies such asomalizumab (Xolair®), nucleoside reverse transcriptase inhibitors suchas zidovudine (Retrovir®), abacavir (Ziagen®), abacavir/lamivudine(Epzicom®), abacavir/lamivudine/zidovudine (Trizivir®), didanosine(Videx®), emtricitabine (Emtriva®), lamivudine (Epivir®),lamivudine/zidovudine (Combivir®), stavudine (Zerit®), and zalcitabine(Hivid®), non-nucleoside reverse transcriptase inhibitors such asdelavirdine (Rescriptor®), efavirenz (Sustiva®), nevairapine (Viramune®)and etravirine (Intelence®), nucleotide reverse transcriptase inhibitorssuch as tenofovir (Viread®), protease inhibitors such as amprenavir(Agenerase®), atazanavir (Reyataz®), darunavir (Prezista®),fosamprenavir (Lexiva®), indinavir (Crixivan®), lopinavir and ritonavir(Kaletra®), nelfinavir (Viracept®), ritonavir (Norvir®), saquinavir(Fortovase® or Invirase®), and tipranavir (Aptivus®), entry inhibitorssuch as enfuvirtide (Fuzeon®) and maraviroc (Selzentry®), integraseinhibitors such as raltegravir (Isentress®), doxorubicin(Hydrodaunorubicin®), vincristine (Oncovin®), bortezomib (Velcade®),and/or dexamethasone (Decadron®) in combination with lenalidomide(Revlimid®), or any combination(s) thereof.

In another embodiment, the present invention provides a method ofpreventing and/or treating gout comprising administering to a patient inneed thereof a compound of formula I and one or more additional activeagents selected from non-steroidal anti-inflammatory drugs (NSAIDS) suchas aspirin, ibuprofen, naproxen, etodolac (Lodine®) and celecoxib,colchicine (Colcrys®), corticosteroids such as prednisone, prednisolone,methylprednisolone, hydrocortisone, and/or the like, probenecid,allopurinol and febuxostat (Uloric®).

In another embodiment, the present invention provides a method ofpreventing and/or treating rheumatoid arthritis comprising administeringto a patient in need thereof a compound of formula I and one or moreadditional therapeutic agents selected from non-steroidalanti-inflammatory drugs (NSAIDS) such as but not limited to aspirin,ibuprofen, naproxen, etodolac (Lodine®) and celecoxib, corticosteroidssuch as prednisone, prednisolone, methylprednisolone, hydrocortisone,and the like, sulfasalazine (Azulfidine®), antimalarials such ashydroxychloroquine (Plaquenil®) and chloroquine (Aralen®), methotrexate(Rheumatrex®), gold salts such as gold thioglucose (Solganal®), goldthiomalate (Myochrysine®) and auranofin (Ridaura®), D-penicillamine(Depen® or Cuprimine®), azathioprine (Imuran®), cyclophosphamide(Cytoxan®), chlorambucil (Leukeran®), cyclosporine (Sandimmune®),leflunomide (Araya®) and “anti-TNF” agents such as etanercept (Enbrel®),infliximab (Remicade®), golimumab (Simponi®), certolizumab pegol(Cimzia®) and adalimumab (Humira®), “anti-IL-1” agents such as anakinra(Kineret®) and rilonacept (Arcalyst®), antibodies such as rituximab(Rituxan®), “anti-T-cell” agents such as abatacept (Orencia®) and/or“anti-IL-6” agents such as tocilizumab (Actemra®).

In some embodiments, the present invention provides a method ofpreventing and/or treating osteoarthritis comprising administering to apatient in need thereof a compound of formula I and one or moreadditional therapeutic agents selected from acetaminophen, non-steroidalanti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen,etodolac (Lodine®) and celecoxib, diclofenac, cortisone, hyaluronic acid(Synvisc® or Hyalgan®), and/or monoclonal antibodies such as tanezumab.

In some embodiments, the present invention provides a method ofpreventing and/or treating lupus comprising administering to a patientin need thereof a compound of formula I and one or more additionaltherapeutic agents selected from acetaminophen, non-steroidalanti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen,etodolac (Lodine®) and celecoxib, corticosteroids such as prednisone,prednisolone, methylprednisolone, hydrocortisone, and the like,antimalarials such as hydroxychloroquine (Plaquenil®) and chloroquine(Aralen®), cyclophosphamide (Cytoxan®), methotrexate (Rheumatrex®),azathioprine (Imuran®) and/or anticoagulants such as heparin(Calcinparine® or Liquaemin®) and warfarin (Coumadin®).

In some embodiments, the present invention provides a method ofpreventing and/or treating inflammatory bowel disease comprisingadministering to a patient in need thereof a compound of formula I andone or more additional therapeutic agents selected from mesalamine(Asacol®) sulfasalazine (Azulfidine®), antidiarrheals such asdiphenoxylate (Lomotil®) and loperamide (Imodium®), bile acid bindingagents such as cholestyramine, alosetron (Lotronex®), lubiprostone(Amitiza®), laxatives such as Milk of Magnesia, polyethylene glycol(MiraLax®), Dulcolax®, Correctol® and Senokot® and anticholinergics orantispasmodics such as dicyclomine (Bentyl®), anti-TNF therapies,steroids, and/or antibiotics such as Flagyl or ciprofloxacin.

In some embodiments, the present invention provides a method ofpreventing and/or treating asthma comprising administering to a patientin need thereof a compound of formula I and one or more additionaltherapeutic agents selected from Singulair®, beta-2 agonists such asalbuterol (Ventolin® HFA, Proventil® HFA), levalbuterol (Xopenex®),metaproterenol (Alupent®), pirbuterol acetate (Maxair®), terbutalinesulfate (Brethaire®), salmeterol xinafoate (Serevent®) and formoterol(Foradil®), anticholinergic agents such as ipratropium bromide(Atrovent®) and tiotropium (Spiriva®), inhaled corticosteroids such asprednisone, prednisolone, beclomethasone dipropionate (Beclovent®,Qvar®, and Vanceril®), triamcinolone acetonide (Azmacort®), mometasone(Asthmanex®), budesonide (Pulmocort®), flunisolide (Aerobid®), Afviar®,Symbicort®, and Dulera®, cromolyn sodium (Intal®), methylxanthines suchas theophylline (Theo-Dur®, Theolair®, Uniphyl®, Theo-24®) andaminophylline, and/or IgE antibodies such as omalizumab (Xolair®).

In some embodiments, the present invention provides a method ofpreventing and/or treating COPD comprising administering to a patient inneed thereof a compound of formula I and one or more additionaltherapeutic agents selected from beta-2 agonists such as albuterol(Ventolin® HFA, Proventil® HFA), levalbuterol (Xopenex®), metaproterenol(Alupent®), pirbuterol acetate (Maxair®), terbutaline sulfate(Brethaire®), salmeterol xinafoate (Serevent®) and formoterol(Foradil®), anticholinergic agents such as ipratropium bromide(Atrovent®) and tiotropium (Spiriva®), methylxanthines such astheophylline (Theo-Dur®, Theolair®, Uniphyl®, Theo-24®) andaminophylline, inhaled corticosteroids such as prednisone, prednisolone,beclomethasone dipropionate (Beclovent®, Qvar®, and Vanceril®),triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®), budesonide(Pulmocort®), flunisolide (Aerobid®), Afviar®, Symbicort®, and/orDulera®.

Addition embodiments of the present invention are described below. Theseembodiments are illustrative and should not be construed as limiting thescope of the claimed invention.

Embodiment 1. A pharmaceutical composition for preventing and/ortreating inflammatory diseases, the composition comprising, as an activesubstance, a compound represented by Chemical Formula 1, or an isomer orpharmaceutically acceptable salt thereof:

(where, in Chemical Formula 1 above,R¹ is C₁-C₃ alkoxy;R² and R³ are each independently hydrogen, straight chain or branchedchain C₁-C₁₀ alkyl, or C₃-C₆ cycloalkyl; andR⁴ is haloalkyl).

Embodiment 2. The pharmaceutical composition for preventing and/ortreating inflammatory disease of embodiment 1, the compositioncomprising, as an active substance, a compound wherein, R¹ is methoxy;R² is straight chain or branched chain C₁-C₅ alkyl, or C₃-C₄ cycloalkyl;R³ is hydrogen; and R⁴ may be trifluoromethyl, or an isomer orpharmaceutically acceptable salt thereof.

Embodiment 3. The pharmaceutical composition for preventing and/ortreating inflammatory disease of embodiment 1, the compositioncomprising, as an active substance, a compound, or an isomer orpharmaceutically acceptable salt thereof, where the compound representedby Chemical Formula 1 may be(4-((−4-ethylamino)-3-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-6-yl)amino)-3-methoxyphenyl)(4-morpholinopiperidine-1-yl)methanone.

Embodiment 4. The pharmaceutical composition for preventing and/ortreating inflammatory disease of any one of embodiment 1 throughembodiment 3, the composition comprising, as an active substance, acompound, or an isomer or pharmaceutically acceptable salt thereof,where the inflammatory disease is at least one selected from a groupcomprising asthma, acute lung injury, graft versus host disease (GvHD),chronic obstructive pulmonary disease, allergy, systemic lupuserythematosus, scleroderma, inflammatory bowel disease (for example,ulcerative colitis and Crohn's disease), atopic dermatitis, psoriasis,anaphylaxis, dermatitis, diabetic retinosis, retinitis, maculardegeneration, uveitis, conjunctivitis, rheumatoid arthritis, ankylosingspondylitis, osteoarthritis, osteoporosis, diabetes, diabetic kidneydisease, nephritis, Sjogren's syndrome, Crohn's disease, autoimmunepancreatitis, periodontal disease, alopecia areata, graft versus hostdisease, chronic pelvic inflammatory disease, endometriosis, rhinitis,tonsilitis, otitis media, sore throat, cystitis and chronic prostatitis.

Embodiment 5. The pharmaceutical composition for preventing and/ortreating inflammatory disease of any one of embodiment 1 throughembodiment 3, the composition comprising, as an active substance, acompound, or an isomer or pharmaceutically acceptable salt thereof,where the inflammatory disease is at least one selected from a groupcomprising inflammatory bowel disease, rheumatoid arthritis, lupus,psoriasis, atopic dermatitis, graft versus host disease (GvHD), acutelung injury, alopecia areata, and osteoarthritis.

Embodiment 6. The pharmaceutical composition for preventing and/ortreating inflammatory disease of any one of embodiment 1 throughembodiment 3, the composition comprising, as an active substance, acompound, or an isomer or pharmaceutically acceptable salt thereof,which inhibits protein kinase activity.

Embodiment 7. The pharmaceutical composition for preventing and/ortreating inflammatory disease of any one of embodiment 1 throughembodiment 3, the composition comprising, as an active substance, acompound, or an isomer or pharmaceutically acceptable salt thereof,which inhibits inflammation-related factors.

Hereinafter, the present invention will be described in further detailthrough examples and experimental examples.

However, the following examples and experimental examples are intendedonly to exemplify the present invention, and the scope of the presentinvention is not limited to the following examples and experimentalexamples.

EXEMPLIFICATION General Synthetic Methods

The compounds of the present invention were prepared according to themethods described in Korean Registered Patent Gazette No. 10-1896568.Chemical structures, compound names, and H¹ NMR data for compounds 1 to5 are showed in Table 1, below.

TABLE 1 Cmpd Compound structure Compound name ¹H NMR; MS(ESI) m/z 1

(4-((4-(ethylamino)- 3-(trifluoromethyl)- 1H-pyrrolo[2,3- b]pyridine-6-yl)amino)-3- methoxyphenyl)(4- morpholinopiperidine- 1-yl)methanone ¹HNMR (400 MHz, HCl salt, DMSO) δ 11.99 (s, 1H), 11.29 (s, 1H), 9.01 (brs, 1H), 8.20 (br s, 1H), 7.59 (s, 1H), 7.11 (s, 1H), 7.02 (d, J = 8.1Hz, 1H), 6.12 (s, 1H), 5.41 (br s, 1H), 4.59-3.91 (m, 4H), 3.89 (s, 3H),3.87- 3.80 (m, 2H), 3.38-3.19 (m, 5H), 3.15-2.79 (m, 4H), 2.25-2.12 (m,2H), 1.76-1.66 (m, 2H), 1.25 (d, J) = 7.1 Hz, 3H); 547 [M + H]⁺ 2

(3-methoxy-4-((4- (methylamino)-3- (trifluoromethyl)- 1H-pyrrolo[2,3-b]pyridine-6- yl)amino)phenyl)(4- morpholinopiperidine- 1-yl)methanone¹H NMR (400 MHz, TFA, salt, Methanol-d₄) δ 7.55 7.46 (m, 2H), 7.23 (d, J= 1.4 Hz, 1H), 7.13 (dd, J = 8.0, 1.5 Hz, 1H), 5.94 (s, 1H), 4.17 4.00(m, 2H), 3.93 (s, 3H), 3.87 3.75 (m, 2H), 3.63 3.44 (m, 4H), 3.30 3.15(m, 4H), 3.04 (s, 3H) 3.02 2.85 (m, 1H), 2.36 2.13 (m, 2H), 1.85 1.70(m, 2H); 533 [M + H]⁺ 3

(4-(4- (isopropylamino)-3- (trifluoromethyl)- 1H-pyrrolo[2,3-b]pyridine-6- ylamino)-3- methoxyphenyl)(4- morpholinopiperidine-1-yl)methanone ¹H NMR (400 MHz, TFA salt, MeOD- d₄) δ 7.54-7.52 (m, 2H),7.23 (s, 1H), 7.14 (d, J = 8.04 Hz, 1H), 5.98 (s, 1H), 4.09-3.98 (m,3H), 3.93 (s, 3H), 3.89-3.84 (m, 3H), 3.55-3.40 (m, 4H), 3.33-3.13 (m,4H), 2.24-2.15 (m, 2H), 1.77-1.74 (m, 2H), 1.35 (d, J = 6.28 Hz, 6H);561 [M + H]⁺ 4

(S)-(4-((4-(2- butylamino)-3- (trifluoromethyl)- 1H-pyrrolo[2,3-b]pyridin-6- yl)amino)-3- methoxyphenyl)(4- morpholinopiperidine-1-yl)methanone 1H NMR (400 MHz, TFA salt, MeOD) δ 7.41 (s, 1H), 7.40 (d,J = 8.0 Hz, 1H), 7.12 (s, 1H), 6.75 (d, J = 8.0 Hz, 1H), 5.86 (s, 1H),4.08-3.96 (m, 2H), 3.82 (s, 3H), 3.79-3.51 (m, 3H), 3.49-3.35 (m, 3H),3.29-3.04 (m, 4H), 2.25-2.04 (m, 2H), 1.84-1.63 (m, 4H), 1.27-1.18 (m,4H), 0.96-0.84 (m, 4H); 575 [M + H]⁺ 5

(4-((4- (cyclopropylamino)- 3- (trifluoromethyl)-1- ((2-(trimethylsilyl) ethoxy)methyl)-1H- pyrrolo[2,3-b] pyridine-6-yl) 3-methoxyphenyl) (4- morpholinopiperidine- 1-yl)methanone ¹H NMR (400 MHz,TFA salt, MeOD- d₄) δ 7.62 (d, J = 8.18 Hz, 1H), 7.50 (s, 1H), 7.24 (s,1H), 7.15 (d, J = 7.27 Hz), 1H), 6.42 (s, 1H), 4.10-3.99 (m, 2H),3.98-3.94 (m, 1H), 3.94 (s, 3H), 3.93-3.79 (m, 2H), 3.72-3.49 (m), 3H),3.34-3.13 (m, 5H), 2.70-2.65 (m, 1H), 2.40-2.14 (m, 2H), 1.85-1.60 (m,2H), 1.00-0.94 (m, 2H), 0.76-0.68 (m, 2H); 559 [M + H]⁺

Example 1. Evaluation of Inhibitory Activity of the Compounds Accordingto the Present Invention on Various Kinases

Measuring the enzyme (kinase) selectivity and inhibitory activity ofCompound 1 selected from among the example compound of the presentinvention was consigned to DiscoverX, and a scanMAX™ Kinase analysispanel was used to carry out the experiment. Here, the concentration ofthe drug used to treat the enzyme was 1 μM in DMSO; the percentagecontrol (% control) was determined using the following method, and theresults are shown in Table 2.

[(Example compound−positive control)/(negative control−positivecontrol)×100]

Here, the positive control refers to a compound showing a percentagecontrol of 0%, and the negative control indicates a percentage controlof 100% with DMSO. Furthermore, regarding the enzyme selectivity of thepresent invention, if the percentage control for each enzyme was <41%(that is, less than 41%), the compound was judged to have activity withregard to such enzyme.

TABLE 2 Protein Protein Item kinase Item kinase Protein ALKALK(C1156Y)Protein ABL1- kinases ALK(L1196M) kinases nonphosphorylated exhibitingCLK1 exhibiting CAMK2D inhibition CLK2 inhibition CAMKK2 of 59% or CLK3of at least CHEK2 more CLK4 40% and up CSNK1A1 CSNK1D to 59% CSNK1EDYRK1A ERK5 DYRK1B HUNK DYRK2 INSR GAK JAK1(JH2domain- JNK1pseudokinase) LRRK2(G2019S) JNK2 LTK JNK3 MYLK LRRK2 PAK2 MAPKAPK2 PHKG1PLK4 PHKG2 STK39 STK33

As shown in Table 2 above, Compound 1 exhibits inhibitory activityagainst various protein kinases.

Example 2. Evaluation of Pro-Inflammatory Cytokine Inhibitory Activityof the Compounds According to the Present Invention

THP-1 cells were pre-treated with Compound 1 for 1 hour, and thentreated with lipopolysaccharides (Lipopolysaccharides, LPS, 0.5 μg/ml),an inflammatory substance, for 24 hours to confirm the expression of thepro-inflammatory cytokines TNF-α and IL-6 by the THP-1 cells and theNF-κb signaling pathway associated with their expression.

When the THP-1 cells were treated with LPS, the expression of theinflammation-inducing pro-inflammatory cytokines TNF-α and IL-6increased, and their expression was effectively inhibited by treatmentwith Compound 1 (FIG. 1a ); the expression of such pro-inflammatorycytokines is increased through activation of the NF-κb signalingpathway, and it can be seen that intranuclear movement of p50 and p65,which are associated with the NF-κb signaling pathway, is effectivelyinhibited by Compound 1 (FIG. 1b ). These results confirm thatexpression of pro-inflammatory cytokines by immune cells is effectivelyinhibited by Compound 1, and can be interpreted as demonstratingpotential for use of Compound 1 in treatment of inflammatory diseases.

Example 3. Evaluation of the Regulation Activity of the CompoundsAccording to the Present Invention Against Regulatory T Cell and Th17Cell Differentiation

Using naive T cells in the mouse spleen, Compound 1 was sufficientlydissolved in DMSO for the differentiation conditions of regulatory Tcells and Th17 cells to confirm differentiation. The experiment wasconducted using Harmine as a control.

In the results, as shown in FIG. 2, when treated with Compound 1, it wasfound that differentiation of regulatory T cells (T_(reg)) increased anddifferentiation of Th17 cells decreased, indicating potential for use intreatment of inflammatory diseases such as autoimmune disease.

Example 4. Evaluation of the Efficacy of the Compound of the PresentInvention Using a Collagen-Induced Arthritis Mouse Model

In the present experiment, male DBA-1 mice aged 8 to 10 weeks were usedas the animal model. After mixing Bovine Type II Collagen (CII) with thesame amount of complete Freud's adjuvant, 100 μL was injected byintradermal injection into the tail of the DBA-1 mouse. After 21 days,the same CII was mixed with the same amount of incomplete Freud'sadjuvant (IFA, Chondrex), then 100 μL was injected by intradermalinjection into the tail. Observation of symptoms of the mouse modelbegan at this point, and arthritis assessment was carried out everyother day. Compound 1 was sufficiently dissolved in a solvent (1% citricacid+20% HP-beta-CD) and then orally administered at 30 mg/kg twice aday for 2 weeks starting on day 29. In the case of the control groupEnbrel (anti-TNF-α), 10 mg/kg was subcutaneously administered once every2 days, then dexamethasone (0.15 mg/kg), Baricitinib (5 mg/kg),Filgotinib (50 mg/kg), and Tofacitinib (10 mg/kg) were orallyadministered twice a day; as for arthritis assessment, the scoresassessed at the four feet of the mouse were added and divided by 4 togive a mean, based on the mean arthritic index by Rosloniec et al(Reference Literature 1: Brand D D, et al. Nat Protoc. 2007; 2(5):1269-75).

The scores and criteria for the arthritis assessment are as follows.

Score 0: No edema or erythema.

Score 1: Mild edema and erythema limited to the foot or ankle joint

Score 2: Mild edema and erythema from the ankle joint to the tarsals

Score 3: Moderate edema and erythema from the ankle joint to themetatarsals.

Score 4: Severe edema and skin flaring from the ankle to the entire leg,or stiffness in the leg

Upon termination of the experiment, the weight of the two hind legs ofeach mouse were measured and added, and these measurements werecompared.

Furthermore, for histological examination of the joint area of thecontrol and the experimental group treated with Compound 1, the tissueswere dyed with H&E and T-blue and observed under an optical microscopeto evaluate the sum of the degree of joint fibrosis due to necrosis andinflammatory cell infiltration (0 to 5), the structure of the cartilageunder examination, cell count and distribution, synovial inflammationand hyperplasia (Modified Rankin Scores, 0 to 12) according to thecriteria of Table 3 and the results were compared (Reference 2: CynthiaShackelford, et al. Toxicologic Pathology. Vol. 30, No. I, p93-96;Reference 3: Pine P R, et al. Clin Immunol. 124 (3): 244-57).

TABLE 3 Arthritis fibromatous with necrosis Modified Rankin Scores andSynovial inflammatory inflammation cell and Score infiltration StructureCells hyperplasia 0 Normal Normal Normal Normal 1 Less than 1% IrregularBroad Normal or minute surface hyperplasia abnormal growth formationaccompanying mild inflammation 2  1-25% Pannus and Replication Moderateirregular inflammation surface accompanied by formation mildtrophoblastic growth 3 26-50% Splitting at Hypocellular Moderate thechanging state trophoblastic growth surface (necrosis) accompanyingmoderate inflammation 4 51-75% Splitting Moderate of the trophoblasticgrowth tide mark accompanying severe inflammation 5 76-100%  Splittingof the cartilage 6 Complete destruction of cartilage tissue

The results of evaluating the arthritis index after inducinginflammation with collagen are shown in FIG. 3. It was determined in theresults that the increased arthritic index was reduced when the compoundof the present invention was administered, and the weight of the joint,which had increased due to induction of arthritis, was reduced (FIGS. 3ato 3b ). Furthermore, based on photography of the joint area forhistological analysis, it can be seen that treatment with Compound 1reduced the area of cells were collagen-induced inflammation hadoccurred, and also reduced the area of the damaged joint (FIGS. 3c to 3e). This confirms that the above compound inhibits inflammatory reactionand has the potential to treat and suppress the progression ofarthritis.

Example 5. Evaluation of Inflammatory Cytokine Expression InhibitionActivity in Peripheral Blood Mononuclear Cells (PBMC) of InflammatoryBowel Disease Patients

Blood was collected from patients with inflammatory bowel disease andthe peripheral blood mononuclear cells were isolated within 12 hours ofcollection; by treating for 24 hours with Compound 1, inflammatorycytokine secretion inhibition activity in peripheral blood mononuclearcells was examined.

The peripheral blood mononuclear cells of inflammatory bowel diseasepatients are already activated and express inflammatory cytokines. Itwas confirmed by treating the peripheral blood mononuclear cells ofpatients with inflammatory bowel disease with Compound 1 effectivelyinhibiting expression of the inflammatory cytokines TNF-α, IL-6 andIL-17A, and this result can be interpreted as indicating potential ofthe compound for use in treatment of inflammatory bowel disease (FIG.4).

Example 6. Evaluation of the Efficacy of Compounds Using a2,4,6-Trinitrobenzenesulfonic Acid (TNBS) Induced Colitis Mouse Model

In the present experiment, male BALB/c mice (8 weeks old) were used. Theexperimental animals were subjected to dietary restriction for one dayprior to treating with TNBS. Bowel inflammation was induced byadministering TNBS (TNBS solution, 1 mg in 0.1 mL 50% ethanol) bysubmucosal injection in the colon. For the control animals in whichinflammation was not induced, only 0.9% NaCl solution without TNBS wasadministered by submucosal injection in the colon.

Compound 1 was sufficiently dissolved in a solvent (5% DMSO, 5% PEG400,90% DDW within 1% Tween 80) and then orally administered once daily atthe prescribed concentration (60 mpk). The comparative compound,cyclosporine A, was orally administered once daily, and the TNF-αblocker (Enbrel®) was administered intraperitoneally once daily.

During the test, changes in body weight, occult bleeding in stool, andstool consistently were examined every day, and serum was collected onday 5 of the experiment. Bowel tissue was collected at the terminationof the experiment to measure the weight of the bowel tissue, degree ofpolymorphonuclear leukocyte (PMN) deposition and degree ofmyeloperoxidase (MPO) expression.

In the results, as shown in FIG. 6, MPO levels were lower than the MPOlevels of the control not treated with the compound. Furthermore,weighing the bowel tissue showed a weight reduction in the experimentalgroup treated with Compound 1, and this can be interpreted as being dueto the reduction of the edema, etc. caused by inflammation.Additionally, the inflammation sites on the collected bowel tissue weredyed with H&E and photographed with an optical microscope, and theresults showed that the lamina propria, whose thickness had increaseddue to induction of inflammation, decreased in thickness due totreatment with the compound of the present invention (FIG. 5).

Example 7. Evaluation of the Efficacy of the Compounds Using aDSS-Induced Inflammatory Bowel Disease Mouse Model

The animals used for the present experiment was male C57BL/6 mice (aged6 to 8 weeks) weighting 20 to 24 g, purchased from Charles RiverLaboratories. All the mice except the normal control were made to drink3% DSS water for 5 days to induce enteritis.

Starting the day after induction of enteritis began, Compound 1 wassufficiently dissolved in solvent (0.5% citric acid in dH₂0) then orallyadministered twice a day at a prescribed concentration (20 mg/kg). Thecomparative compound cyclosporine A was dissolved in 0.5%methylcellulose solvent and orally administered once a day at aprescribed concentration (20 mg/kg), and Tofacitinib was dissolved in0.5% methylcellulose solvent and orally administered twice a day at aprescribed concentration (10 mg/kg).

Changes in body weight, occult bleeding in stool, and stool consistentlywere examined daily during the experiment to assess the degree ofdisease activity, and upon termination of the experiment (day 12), boweltissue was collected in order to measure bowel tissue weight and length,perform histological analysis using H&E dyeing and PAS dyeing, andanalyze expression of inflammatory cytokines in the bowel tissue.

To assess disease activity, changes in weight and other criteriadisclosed in Table 4 below were used for scoring; scores wereaggregated.

TABLE 4 Weight Stool Occult blood Score loss consistency test Activity 0 0.0-4.99% Normal None Active 1  5.0-9.99% 2 10.0-14.99% SlightlyUrinary occult blood Obtusion watery stool and slightly fecal occultblood 3 15.0-19.99% 4   >20.0% Diarrhea Severe fecal occult No movementblood without stimulation 5   >30.0% Dying

In the results, as shown in FIG. 7, treatment with the compound showedless weight reduction compared to the control group not treated with thecompound, and disease activity was reduced to the level of the controladministered cyclosporine A. Furthermore, as shown in FIG. 8, whereasthe length to weight ratio of bowel tissue increases as colitis becomesmore severe, administration of Compound 1 reduced the bowel tissuelength to weight ratio, and histological analysis confirmed that loss ofgoblet cells was also reduced. Additionally, it was confirmed thatadministration of Compound 1 decreased the expression of inflammatorycytokines (TNR-α, IL-6, IL-17A), which increases as inflammation inbowel tissue is aggravated. In summation, as Compound 1 was confirmed toreduce symptoms caused by colitis (diarrhea, fecal occult bleeding, andinflammatory markers), the results can be interpreted to confirm itspotential for use in treating colitis.

Example 8. Evaluation of the Efficacy of the Compounds Using a HouseDust Mite-Induced Atopic Dermatitis Model

The backs and ears of NC/Nga mice aged 8 weeks were shaved, and 0.1 ghouse dust mite cream was applied once every three days for a total ofseven times to induce atopic dermatitis. For the negative control groupwhere dermatitis was not induced, a cream not including house dust miteswas applied. After inducing atopic dermatitis, Compound 1 wassufficiently dissolved in solvent (0.5% citric acid), then orallyadministered twice daily for 14 days at prescribed concentrations,respectively 10, 20 and 30 mpk. To the comparative control group, theJAK1 inhibitor upadacitinib was orally administered at 6 mg/kg twicedaily for 14 days.

After inducing atopic dermatitis using house dust mite cream, symptomsas well as the state of the skin of the ears and backs were scored twiceweekly starting from the initial date of administration; themeasurements were added and the means are shown in FIG. 9. Furthermore,on the date of termination of the experiment, skin tissue and serum werecollected to observe the level of expression of inflammatory cytokinesin the tissue and the level of expression of IgE in serum using ELISAanalysis; the results are shown in FIG. 10.

In the results, dermatitis on the back and ears was alleviated in aconcentration-dependent manner for administration of Compound 1, andadministration at 30 mpk was most effective at alleviating dermatitis;it was confirmed that Compound 1 was more effective at alleviatingatopic dermatitis than the upadacitinib control group. Aggravation ofatopic dermatitis leads to increased inflammatory cytokines that induceinflammation in the lesion area, and comparing the degree ofinflammatory cytokine expression in the back tissue in the groupadministered Compound 1 at 30 mpk against the control group, astatistically significant reduction was confirmed. Furthermore, it isreported that IgE in serum increases if atopic dermatitis is induced ina house dust mite cream-induced atopic dermatitis model, and in thegroup administered Compound 1, a concentration-dependent reduction inIgE expression was confirmed, with a statistically significant reductionobserved in the group administered at 30 mpk. From the above results, itcan be confirmed that Compound 1 has a therapeutic effect on atopicdermatitis.

Example 9. Evaluation of In-Vivo Stability and Bioavailability of theCompounds According to the Present Invention

To evaluate the in-vivo stability and bioavailability of Compound 1according to the present invention for use as a drug, metabolicstability evaluation, a metabolic enzyme CYP inhibition assay, hERGanalysis, CaCo-2 analysis to observe biomembrane permeability of thedrug, and plasma stability evaluation were carried out. The results areshown in Table 5 below. MS is an index of in-vivo metabolic stability,and values confirmed that Compound 1 is safe in mice up to dogs, whichare larger animals.

DDI (Drug Interaction) can be observed in a CYP inhibition assay. Aluminescence assay was used to measure and evaluate inhibition activityagainst 1A2, 2C9, 2C19, 2D6 and 3A4, which are important Phase 1 drugmetabolism CYP450 enzymes involved in drug metabolism. Whereas the CYP3A4 index is normally suppressed, almost no enzyme inhibition activitywas exhibited by the compound of the present invention.

The hERG analysis is used as an indicator of cardiac toxicity, andgenerally a value of 5 μM indicates safety; the compound of the presentinvention was confirmed to have excellent safety in terms of cardiactoxicity.

Caco-2 permeability observes the cell membrane permeability of a drug,and can be considered an index associated with drug absorption andexcretion. An ER ratio value close to “1” indicates almost equivalentdrug absorption and excretion, and the compound of the presentinvention, exhibiting a value of 1.08, was confirmed to have excellentcell membrane permeability.

Hydrolysis in plasma is a factor which, together with metabolicreactions, impacts fast in-vivo decomposition of a drug and shorthalf-life thereof; plasma stability is a test which assesses these. Thecompound at a certain concentration was added to plasma and reacted fora certain period, then collected, and LC-MS/MS was used to measure thepercentage of compound remaining relative to the amount prior to thereaction.

Even after 2 hours, 97.2% of the compound of the present inventionremained in plasma without being degraded, and thus exhibited excellentplasma stability.

TABLE 5 Plasma hERG Caco-2 stability MS (1 μM) (% remaining during 30min) CYP (10 μM) (% of control activity IC50 permeability (% remaining,Cmpd Human Mouse Rat Dog Monkey 1A2 2C9 2C19 2D6 3A4 (μM) (ER ratio) 30and 120 min 1 39 40 20 96 — 86 74 84 92 72 4.39 0.201 100, 97.2 (1.08)

Example 10. Evaluation of Topical Anti-Inflammatory Activity of Compound1 in the Imiquimod (IMQ)-Induced Psoriasis-Like Skin Inflammation

Compound 1 (0.3%, 1% and 3%) and the vehicle (5% DMSO/75% PEG400/20%EtOH) were topically administered at 20 μL/mouse on the right ear oncedaily (QD) for 9 consecutive days. Dexamethasone (0.15%), the referencecompound, was applied topically at 20 μL/mouse on the right ear oncedaily (QD) during the same study period.

Fifteen (15) mg imiquimod (IMQ) cream (5%) (Aldara; 3M Pharmaceuticals)was topically administered once daily (QD) on the right ear one hourafter treatment from Day 1 to Day 9 for 9 consecutive days, translatingto a daily dose of 0.75 mg of the active compound.

Ear swelling was measured on Day 0 and thereafter 30 min before dosingon Days 2, 4, 6 and 8. On Day 10, ear swelling was measured 24 hoursafter the last dosing.

The results are shown in FIG. 11a and FIG. 11b . Topical application ofmouse ear with 5% IMQ cream, a TLR7/8 ligand and potent immuneactivator, triggered a psoriasis-like inflammation and showed asignificant (p<0.05) increase in ear swelling compared to the shamcontrol, indicating a successful induction of IMQ-induced psoriasis inno treatment group. However, topical administration of vehicle control(5% DMSO/75% PEG400/20% EtOH) was associated with a significant (p<0.05)reduction of IMQ-induced ear swelling from Day 6 to Day 10, whencompared to the no treatment group.

Compound 1 (0.3, 1% and 3%) had moderate to significant (p<0.05)inhibition on IMQ-induced ear swelling from Day 6 to Day 10 in adose-dependent manner, relative to the no treatment and vehicle controlgroups. The reference compound, dexamethasone (0.15%), significantly(p<0.05) reduced the IMQ-induced ear swelling in BALB/c mice.

No significant differences in the thickness of left ear were observedbetween no treatment and vehicle control groups, but topicaladministrations of dexamethasone (0.15%) also caused significant(p<0.05) decreases in left ear swelling during the study period. The earweight measurements were consistent with the net swelling measurements.Body weight gain between the vehicle control and treated groups weresimilar, only dexamethasone showed significant (p<0.05) decrease in bodyweight during the study period.

Per histopathological examination of the ear skin, epidermalhyperplasia, multifocal to diffuse inflammatory cell infiltration,multifocal necrosis and epidermal hyperkeratosis lesions were observedin the no treatment group and vehicle control groups. The severity ofskin lesions was attenuated significantly by dexamethasone (0.15%), andCompound 1 (3%), when compared to the no treatment group and vehiclegroup.

In conclusion, Compound 1 (3%, high dose) at 20 μL/mouse QD×9 days giventopically had significant (p<0.05) protective effects in IMQ-inducedpsoriasis-like inflammation, as evidenced by the improvements in earswelling, ear weight and histopathological result in IMQ inducedpsoriasis model in BALB/c mice.

Example 11. Evaluation of Oral Anti-Inflammatory Activity of theCompounds in the Imiquimod (IMQ)-Induced Psoriasis-Like SkinInflammation

Compound 1 at 45 and 60 mg/kg and the vehicle (0.5% citric acid+20%HP-b-CD in DDW) at 5 mL/kg were given by oral gavage twice (BID) dailyfor 9 consecutive days. The positive control, methotrexate at 2 mg/kgand tofacitinib at 5 mg/kg were administered by oral gavage twice (BID)daily during the same study period.

Fifteen (15) mg Imiquimod (IMQ) cream (5%) (Aldara; 3M Pharmaceuticals)was topically (TOP) administered once daily (QD) on the right ear onehour after treatment from Day 1 to Day 9 for 9 consecutive days,translating to a daily dose of 0.75 mg of the active compound. Earswelling was measured on Day 0 and thereafter 30 min before dosing onDays 2, 4, 6 and 8. On Day 10, ear swelling was measured 24 hours afterthe last dosing.

The results are shown in FIG. 12a and FIG. 12b . Topical application onmouse ear with 5% IMQ cream, a TLR7/8 ligand and potent immuneactivator, triggered a psoriasis-like inflammation and showed asignificant (p<0.05) increase in ear swelling compared to the shamcontrol, indicating a successful induction of IMQ-induced psoriasis inboth no treatment and vehicle control (0.5% citric acid+20% HP-b-CD inDDW) groups.

Compound 1 given at 45 and 60 mg/kg had a statistically (p<0.05)significant inhibition on IMQ-induced ear swelling from Day 8 to Day 10,when compared to the vehicle group.

The positive control, methotrexate at 2 mg/kg showed a significant(p<0.05) effect on the IMQ-induced ear swelling on Day 8, when comparedto the vehicle group. Tofacitinib given at 5 mg/kg also showed asignificant (p<0.05) inhibition on IMQ-induced ear swelling from Day 8to Day 10 in the study.

All treated groups showed a little, but significant difference in thethickness of left ear, when compared to the vehicle group during thestudy period. The ear weight measurements were consistent with the netswelling measurements.

Body weight gain between the vehicle control and treated groups werecomparable during the study period. However, methotrexate at 2 mg/kg POBID×9 days significantly decreased the body weight from Day 9 to Day 10in the study.

In conclusion, Compound 1 at 45 and 60 mg/kg PO BID×9 days hadsignificantly (p<0.05) protective effects on IMQ-induced psoriasis-likeinflammation, as evidenced by the improvement in ear swelling in IMQinduced psoriasis model in BALB/c mice.

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments that utilize the compounds and methods of this invention.Therefore, the scope of this invention is to be defined by the appendedclaims rather than by the specific embodiments that have beenrepresented by way of example.

1. A method of preventing or treating an inflammatory disease in a patient in need thereof, the method comprising administering to the patient a compound represented by Chemical Formula 1:

or an isomer, pharmaceutically acceptable salt, or pharmaceutical composition thereof, wherein: R¹ is C₁-C₃ alkoxy; R² and R³ are each independently hydrogen, straight chain or branched chain C₁-C₁₀ alkyl, or C₃-C₆ cycloalkyl; and R⁴ is haloalkyl.
 2. The method of claim 1, wherein the method of preventing or treating an inflammatory disease comprises one or more of: (a) reducing pro-inflammatory cytokines in the patient; (b) increasing differentiation of regulatory T cells (T_(reg)) in the patient; and (c) decreasing pro-inflammatory T cells in the patient.
 3. A method of reducing pro-inflammatory cytokines in a patient in need thereof, the method comprising administering to the patient a compound represented by Chemical Formula 1:

or an isomer, pharmaceutically acceptable salt, or pharmaceutical composition thereof, wherein: R¹ is C₁-C₃ alkoxy; R² and R³ are each independently hydrogen, straight chain or branched chain C₁-C₁₀ alkyl, or C₃-C₆ cycloalkyl; and R⁴ is haloalkyl.
 4. The method of claim 2, wherein the pro-inflammatory cytokine is selected from one or more of NO, IFN-γ, IL-1α, IL-1β, IL-4, IL-6, IL-10, IL-12, IL-13, IL-17, IL-17A, IL-22, IL-23, KC, MIP-1a, MIP-1b, GM-CSF, and TNF-α.
 5. A method of increasing differentiation of regulatory T cells (T_(reg)) in a patient in need thereof, the method comprising administering to the patient a compound represented by Chemical Formula 1:

or an isomer, pharmaceutically acceptable salt, or pharmaceutical composition thereof, wherein: R¹ is C₁-C₃ alkoxy; R² and R³ are each independently hydrogen, straight chain or branched chain C₁-C₁₀ alkyl, or C₃-C₆ cycloalkyl; and R⁴ is haloalkyl.
 6. A method of decreasing pro-inflammatory T cells in a patient in need thereof, the method comprising administering to the patient a compound represented by Chemical Formula 1:

or an isomer, pharmaceutically acceptable salt, or pharmaceutical composition thereof, wherein: R¹ is C₁-C₃ alkoxy; R² and R³ are each independently hydrogen, straight chain or branched chain C₁-C₁₀ alkyl, or C₃-C₆ cycloalkyl; and R⁴ is haloalkyl.
 7. The method of claim 2, wherein the pro-inflammatory T cells are Th17.
 8. The method of claim 1, wherein R¹ is methoxy.
 9. The method of claim 1, wherein, R² is a straight chain or branched chain C₁-C₅ alkyl.
 10. The method of claim 1, wherein, R² is a C₃-C₄ cycloalkyl.
 11. The method of claim 1, wherein, R³ is hydrogen.
 12. The method of claim 1, wherein, R⁴ is trifluoromethyl.
 13. The method of claim 1, wherein the compound represented by Chemical Formula 1 is: (1) (4-((−4-ethylamino)-3-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-6-yl)amino)-3-methoxyphenyl)(4-morpholinopiperidine-1-yl) methanone; (2) (3-methoxy-4-((4-(methylamino)-3-(trifluoromethyl)-1H-pyrrolo[2,3-b] pyridine-6-yl)amino)phenyl)(4-morpholinopiperidine-1-yl)methanone; (3) (4-(4-(isopropylamino)-3-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-6-ylamino)-3-methoxyphenyl)(4-morpholinopiperidine-1-yl)methanone; (4) (S)-(4-((4-(2-butylamino)-3-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)amino)-3-methoxyphenyl)(4-morpholinopiperidine-1-yl)-methanone; or (5) (4-((4-(cyclopropylamino)-3-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b] pyridine-6-yl)3-methoxyphenyl) (4-morpholinopiperidine-1-yl)methanone.
 14. The method of claim 1, wherein the compound represented by Chemical Formula 1 is (4-((−4-ethylamino)-3-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-6-yl)amino)-3-methoxyphenyl)(4-morpholinopiperidine-1-yl) methanone.
 15. The method of claim 1, wherein the inflammatory disease is selected from graft versus host disease (GvHD), obstructive airway disease, chronic obstructive pulmonary disease, allergy, systemic lupus erythematosus, scleroderma, inflammatory bowel disease (e.g., ulcerative colitis and Crohn's disease), atopic dermatitis, psoriasis, anaphylaxis, dermatitis, diabetic retinosis, retinitis, macular degeneration, uveitis, conjunctivitis, rheumatoid arthritis, ankylosing spondylitis, osteoarthritis, osteoporosis, diabetes, diabetic kidney disease, nephritis, Sjogren's syndrome, autoimmune pancreatitis, periodontal disease, alopecia areata, chronic pelvic inflammatory disease, endometriosis, rhinitis, tonsilitis, otitis media, sore throat, cystitis, chronic prostatitis, diseases and conditions of the eye such as ocular allergy, keratoconjunctivitis sicca, vernal conjunctivitis, diseases affecting the nose including allergic rhinitis, and inflammatory disease in which autoimmune reactions are implicated or having an autoimmune component or etiology, including autoimmune hematological disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell anemia and idiopathic thrombocytopenia), polychondritis, Wegener granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Steven-Johnson syndrome, idiopathic sprue, irritable bowel syndrome, celiac disease, periodontitis, hyaline membrane disease, kidney disease, glomerular disease, alcoholic liver disease, multiple sclerosis, endocrine ophthalmopathy, Becker's/Duchenne muscular dystrophy, Grave's disease, sarcoidosis, alveolitis, chronic hypersensitivity pneumonitis, multiple sclerosis, primary biliary cirrhosis, uveitis (anterior and posterior), interstitial lung fibrosis, psoriatic arthritis, systemic juvenile idiopathic arthritis, cryopyrin-associated periodic syndrome, vasculitis, diverticulitis, interstitial cystitis, glomerulonephritis (with and without nephrotic syndrome, e.g. including idiopathic nephrotic syndrome or minimal change nephropathy), chronic granulomatous disease, leptospirosis renal disease, glaucoma, retinal disease, ageing, headache, pain, complex regional pain syndrome, cardiac hypertrophy, muscle atrophy, catabolic disorders, obesity, fetal growth retardation, hypercholesterolemia, heart disease, chronic heart failure, mesothelioma, anhidrotic ectodermal dysplasia, Behcet's disease, incontinentia pigmenti, Paget's disease, pancreatitis, hereditary periodic fever syndrome, asthma (allergic and non-allergic, mild, moderate, severe, bronchitic, and exercise-induced), acute lung injury, acute respiratory distress syndrome, eosinophilia, hypersensitivities, nasal sinusitis, ocular allergy, silica induced diseases, COPD (reduction of damage, airways inflammation, bronchial hyperreactivity, remodeling or disease progression), pulmonary disease, cystic fibrosis, acid-induced lung injury, pulmonary hypertension, polyneuropathy, cataracts, muscle inflammation in conjunction with systemic sclerosis, inclusion body myositis, myasthenia gravis, thyroiditis, Addison's disease, lichen planus, diabetes (Type 1 diabetes or Type 2 diabetes), appendicitis, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic graft rejection, colitis, dacryoadenitis, dermatomyositis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis, fibrositis, gastritis, gastroenteritis, Henoch-Schonlein purpura, hepatitis, hidradenitis suppurativa, immunoglobulin A nephropathy, interstitial lung disease, laryngitis, mastitis, meningitis, myelitis myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis, otitis, parotitis, pericarditis, peritonitis, pharyngitis, pleuritis, phlebitis, pneumonitis, pneumonia, polymyositis, proctitis, prostatitis, pyelonephritis, salpingitis, sinusitis, stomatitis, synovitis, tendonitis, tonsillitis, ulcerative colitis, vaginitis, vasculitis, and vulvitis.
 16. The method of claim 1, wherein the inflammatory disease is selected from inflammatory bowel disease, rheumatoid arthritis, lupus, psoriasis, atopic dermatitis, graft versus host disease (GvHD), acute lung injury, alopecia areata, diabetes, and/or osteoarthritis.
 17. The method of claim 1, wherein the compound represented by Chemical Formula 1 prevents and/or inhibits or otherwise favorably impacts protein kinase activity.
 18. The method of claim 1, wherein the method comprises conjointly administering a compound represented by Chemical Formula 1, or an isomer, pharmaceutically acceptable salt, or pharmaceutical composition thereof and at least one additional active ingredient.
 19. The method of claim 18, wherein at least one additional active ingredient is selected from gamma globulin, phosphodiesterase inhibitors (e.g., apremilast), IRAK4 inhibitors, belimumab, tacrolimus, rapamycin, mycophenolate mofetil, interferon, paracetamol, acetaminophen, non-steroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®), celecoxib, and colchicine (Colcrys®), corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone, betamethasone, and the like, probenecid, allopurinol, febuxostat (Uloric®), sulfasalazine (Azulfidine®), antimalarials such as hydroxychloroquine (Plaquenil®) and chloroquine (Aralen®), methotrexate (Rheumatrex®), gold salts such as gold thioglucose (Solganal®), gold thiomalate (Myochrysine®) and auranofin (Ridaura®), D-penicillamine (Depen® or Cuprimine®), azathioprine (Imuran®), cyclophosphamide (Cytoxan®), chlorambucil (Leukeran®), cyclosporine (Sandimmune®), leflunomide (Araya®) and “anti-TNF” agents such as etanercept (Enbrel®), infliximab (Remicade®), golimumab (Simponi®), certolizumab pegol (Cimzia®) and adalimumab (Humira®), “anti-IL-1” agents such as anakinra (Kineret®) and rilonacept (Arcalyst®), canakinumab (Ilaris®), anti-JAK/STAT inhibitors such as tofacitinib, baricitinib, and GLPG0634, antibodies such as rituximab (Rituxan®), “anti-T-cell” agents such as abatacept (Orencia®), “anti-IL-6” agents such as tocilizumab (Actemra®), diclofenac, cortisone, hyaluronic acid (Synvisc® or Hyalgan®), monoclonal antibodies such as tanezumab, anticoagulants such as heparin (Calcinparine® or Liquaemin®) and warfarin (Coumadin®), antidiarrheals such as diphenoxylate (Lomotil®) and loperamide (Imodium®), bile acid binding agents such as cholestyramine, alosetron (Lotronex®), lubiprostone (Amitiza®), laxatives such as Milk of Magnesia, polyethylene glycol (MiraLax®), Dulcolax®, Correctol® and Senokot®, anticholinergics or antispasmodics such as dicyclomine (Bentyl®), Singulair®, beta-2 agonists such as albuterol (Ventolin® HFA, Proventil® HFA), levalbuterol (Xopenex®), metaproterenol (Alupent®), pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®), salmeterol xinafoate (Serevent®) and formoterol (Foradil®), anticholinergic agents such as ipratropium bromide (Atrovent®) and tiotropium (Spiriva®), inhaled corticosteroids such as beclomethasone dipropionate (Beclovent®, Qvar®, and Vanceril®), triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®), budesonide (Pulmocort®), and flunisolide (Aerobid®), Afviar®, Symbicort®, Dulera®, cromolyn sodium (Intal®), methylxanthines such as theophylline (Theo-Dur®, Theolair®, Slo-Bid®, Uniphyl®, Theo-24®) and aminophylline, IgE antibodies such as omalizumab (Xolair®), nucleoside reverse transcriptase inhibitors such as zidovudine (Retrovir®), abacavir (Ziagen®), abacavir/lamivudine (Epzicom®), abacavir/lamivudine/zidovudine (Trizivir®), didanosine (Videx®), emtricitabine (Emtriva®), lamivudine (Epivir®), lamivudine/zidovudine (Combivir®), stavudine (Zerit®), and zalcitabine (Hivid®), non-nucleoside reverse transcriptase inhibitors such as delavirdine (Rescriptor®), efavirenz (Sustiva®), nevairapine (Viramune®) and etravirine (Intelence®), nucleotide reverse transcriptase inhibitors such as tenofovir (Viread®), protease inhibitors such as amprenavir (Agenerase®), atazanavir (Reyataz®), darunavir (Prezista®), fosamprenavir (Lexiva®), indinavir (Crixivan®), lopinavir and ritonavir (Kaletra®), nelfinavir (Viracept®), ritonavir (Norvir®), saquinavir (Fortovase® or Invirase®), and tipranavir (Aptivus®), entry inhibitors such as enfuvirtide (Fuzeon®) and maraviroc (Selzentry®), integrase inhibitors such as raltegravir (Isentress®), doxorubicin (Hydrodaunorubicin®), vincristine (Oncovin®), bortezomib (Velcade®), and dexamethasone (Decadron®) optionally in combination with lenalidomide (Revlimid®), or any combination(s) thereof.
 20. The pharmaceutical composition of claim 1, further comprising excipients, disintegrating agents, sweetening agents, lubricants, and/or flavoring agents. 